Solid Phase Red Cell Adherence Research Articles

Warm reactive autoantibodies detected by solid phase technology: Patient characteristics, laboratory features, and clinical significance.

This study focused on the serology, clinical characteristics, and hemolytic potential of warm reactive autoantibodies detected by solid phase red cell adherence. Ninety-seven patients with warm autoantibodies were evaluated. Serologic characteristics included the strength of solid phase reactivity, the results of tube-based ancillary testing, direct antiglobulin test and eluate results, and an assessment for contemporaneous alloantibodies. Clinical characteristics of the patients included age, sex, and primary diagnosis. Each patient was also assessed for evidence of hemolysis. Most of the 97 study patients were female (63.9%), and the average age was 66 years. Hematologic disorders were the most common diagnosis. A majority (70.1%) of the warm autoantibodies had 3 to 4+ reaction strengths, and approximately 90% had negative testing with at least 1 test tube method. There was an even distribution of direct antiglobulin test reaction strengths, with 74% reactive with anti-immunoglobulin G only. Alloantibodies were identified in 20% of patients. Evidence of hemolysis was identified in only 13 patients (13.4%). Warm reactive autoantibodies are more likely to be hemolytic, have strongly reactive indirect and direct antiglobulin tests, remain reactive in tube-based ancillary testing methods, and are seen primarily in patients with hematologic disorders.

Comparison of the solid-phase red cell adherence assay and tube method for detection and identification of red blood cell antibodies.

Identifying antibodies to red blood cell antigens is one of transfusion medicine's critical responsibilities. The International Society of Blood Transfusion recognizes 354 red blood cell antigens. Accurate identification of clinically significant alloantibodies is imperative for preventing hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. We compared the performance of the tube (polyethylene glycol-indirect antiglobulin test [PEG-IAT]) and solid-phase red cell adherence assay techniques. We performed a retrospective study on all antibody screens performed between 2007 and 2021 at Stanford Transfusion Services. Initially, 631,535 antibody screens were performed using a solid-phase technique. Subsequent antibody identifications were performed using a combination of tube testing and solid-phase techniques. Antibody screening resulted in 28,316 (4.5%) positive samples. Antibody identification performed on both platforms identified 50 discordant samples. The anti-E antibody had the lowest sensitivity (98.99%) in the automated solid-phase technique, while anti-Jkb had the lowest sensitivity (98.78%) with the PEG-IAT method. To our knowledge, this is the first robust, 15-year study comparing methodologic sensitivity to detect clinically significant alloantibodies. The incidence of discordant results between PEG-IAT and the solid-phase technique was low. Among discordant samples, anti-Jka was commonly detected using the solid-phase method but not with the PEG-IAT. In contrast, anti-E was commonly detected by PEG-IAT but not by the solid-phase method.

[Establish a Graded Method to Avoid HLA Class I Antibodies Corresponding Antigen and Combining HLAMatchmaker Application in Improving the CCI Value after Platelet Transfusion for Patients with IPTR].

To establish a graded method to avoid mean fluorescence intensity (MFI) threshold of HLA Class I antibodies corresponding antigen, and the HLAMatchmaker program has been used to select the minimum mismatch value of donor-patient epitopes. Evaluate the application value of combining both methods in selecting HLA compatible platelets (PTL) for patients with immune platelet transfusion failure (IPTR) in improving platelet the corrected count increment (CCI). A total 7 807 PLT cross-matching compatible were performed by the solid-phase red cell adherence (SPRCA) method for 51 IPTR patients. The Luminex single antigen flow cytometry was used to detect HLA Class I antibodies in patients, and detected the MFI value for different specificity antigens of HLA Class I antibodies, was graded into strong positive group (MFI>4 000, level 1), medium positive group (1 000< MFI≤4 000, 2), weak positive group (500< MFI≤1 000, 3), and one negative control group (MFI≤500). The results of 7 807 SPRCA their negative/positive reaction wells were enrolled and statistically analyzed in different grades and the four groups, the statistical differences between the four groups were compared. Multiple applications for the select HLA Class I compatible donor events were made for patients in two cases, and HLAMatchmaker program was used to calculate the number of HLA Class I epitopes mismatches between the donors and patients. The donor with the minimum number of epitopes mismatches was selected, while avoiding the corresponding antigens of HLA Class I antibodies in levels 1 and 2, the provision of HLA compatible platelets for IPTR. After the transfusions, the CCI value of the platelet transfusion efficacy evaluation index was calculated, and the clinical evaluation of the transfusion effect was obtained through statistical analysis. There were statistically significant differences in the positive results of SPRCA immunoassay among the strong positive group, medium positive group, and weak positive group of 51 IPTR patients with different specific of HLA -I class antibodies and corresponding antigens(all P <0.001). The positive results showed a range from high to low, with strong positive group>medium positive group>weak positive group. There were a statistical difference among between the strongly positive or moderately positive groups and the negative control group(P <0.001). There was no statistical difference between the weakly positive group and the negative control group(P >0.05). The strong positive group was set as the corresponding specific HLA Class I site corresponding antigen grade 1 avoidance threshold, the medium positive group as the grade 2 avoidance thresholds, and the weak positive group as the grade 3 avoidance threshold. In the case of donor platelet shortage, it is not necessary to avoid the weak positive group. Avoiding the strategy of donor antigens and HLAMatchmaker program scores ≤7 corresponding to HLA Class I antibodies of levels 1 and 2, with CCI values>4.5×109/L within 24 hours, can obtain effective clinical platelet transfusion conclusions. When selecting HLA Class I compatible donors for IPTR patients, the grading avoids HLA Class I antibodies corresponding to donor antigens, and the donor selection strategy with the minimum scores of HLAMatchmaker program is comprehensively selected. The negative result confirmed by platelet cross-matching experiments has certain practical application value for improving platelet count in IPTR patients.

Comparison of solid-phase red cell adherence and microcolumn agglutination technology using untreated and enzyme-treated red blood cells.

Screening for clinically significant antibodies is crucial in transfusion medicine and is a routine part of pre-transfusion testing. The indirect antiglobulin test (IAT) is the most reliable and effective test for detecting clinically significant alloantibodies reacting at the antihuman globulin phase. Two of the main methods used for antibody detection and identification are solid-phase red cell adherence (SPRCA) and microcolumn agglutination technology (CAT), with or without enzyme-treated red blood cells (RBCs). This study was undertaken to detect and identify alloantibodies by performing antibody screen (ABS) and antibody identification (ABID) testing using SPRCA and CAT, with and without ficin-treated RBCs. Residual patient samples collected between 1 December 2020 and 19 May 2021 were saved, de-identified, and frozen at ≤-30°C before testing for alloantibodies. Seventy antibodies were detected in 53 samples among the 203 samples that underwent an ABS. Of those samples, 150 (73.0%) were nonreactive, 47 (23.1%) yielded positive results with both CAT and SPRCA, and six (3.0%) yielded positive ABS results with SPRCA only. Fifty-three samples that underwent ABID by both methods yielded eight samples with antibodies identified by SPRCA only. Additional enhancement of the CAT method by the use of ficin-treated RBCs was required to detect seven of the eight SPRCA-only antibodies; one sample remained nonreactive regardless. SPRCA testing detected clinically significant antibodies without the addition of enzyme-treated RBCs that was necessary in the CAT testing.

Evaluation of solid-phase panreactivity with negative direct antiglobulin testing.

Solid-phase red cell adherence (SPRCA) is a sensitive platform for antibody detection, but nonspecific reactions may occur. One pattern of apparent nonspecific reactivity is a panagglutinin with a negative direct antiglobulin test (DAT). The purpose of this study was to define the clinical characteristics of patients with these nonspecific reactions and their associated serologic findings. Twenty patients with panreactive SPRCA testing results were identified between November 2022 and May 2023. In addition to panagglutinins, these patients had (1) a negative polyethylene glycol (PEG) antibody detection test, (2) a negative PEG autocontrol, and (3) a negative DAT. The strength of SPRCA panreactivity and the results of eluate testing (by tube and SPRCA) were studied. Clinical characteristics of patients included age, sex, and primary diagnosis. Each patient was also assessed for evidence of hemolysis. Fourteen female and six male patients were evaluated (average age 44 years). Primary diagnoses included pregnancy (n = 10), acute bleeding (n = 4), orthopedic (n = 3), and other (n = 3). There was no clinical or laboratory evidence of hemolysis. The predominant strength of SPRCA panreactivity was evenly distributed across reaction grades (1+ to 3+). Fifty-five percent of the eluates tested in PEG showed panreactivity, consistent with warm-reactive autoantibodies, while 85 percent of eluates tested by SPRCA were panreactive. Six discrepant cases, in which PEG eluate testing was negative and solid-phase eluate testing showed panreactivity, were associated with weak solid-phase plasma panreactivity (1+). In addition, the reactivity strengths of the eluates tested by SPRCA were invariably more strongly reactive than those eluates tested in PEG. Panagglutination is a distinct SPRCA-only plasma reactivity pattern. Despite a negative PEG tube and DAT, most panagglutinins are warm-reactive autoantibodies. Fortunately, these "interfering" panagglutinins do not appear to be clinically significant and are easily managed by an alternative testing method such as PEG.

The impact of an antibody investigation algorithm emphasizing specificity on reducing potential false-positive warm autoantibody detection at a Canadian tertiary care centre.

To reduce potential false-positive warm autoantibody (WAA) by solid-phase red cell adherence assay (SPRCA), our centre implemented a new antibody investigation algorithm (AIA) by classifying cases with panreactive SPRCA but negative saline-indirect antiglobulin test as 'antibody of undetermined significance' (AUS) after excluding clinically significant antibodies. We assessed the effects of the new AIA and subsequent alloantibody formation in patients with AUS. Samples from patients with positive SPRCA screens between 1 September 2017 and 31 August 2021 were selected for the study. Frequencies of antibodies classified by the old and new AIAs were compared using Fisher's exact test. Patient demographics, transfusion history and antibody formation in cases of AUS were collected. A significant reduction in potential WAA frequencies from 127/1167 (11%) to 53/854 (6%) was observed (p < 0.001) when compared between the old and new AIAs among 2021 positive SPRCA antibody screens. While no patients with AUS later transitioned to potential WAA using the new AIA, four patients developed alloantibodies, including anti-E, anti-C, both anti-C and anti-E, and anti-Wra . A significant reduction in the frequencies of potential false-positive WAA detection at our centre was observed after implementing the new AIA, leading to less resource and phenotypically matched red blood cell (RBC) use. Some patients still developed subsequent RBC alloimmunization, so clinically relevant alloantibodies should be carefully excluded before determining AUS, taking forming or evanescent antibodies into consideration.

Does decreasing the incubation period used in the antibody screen affect its sensitivity?

Pre-transfusion testing (PTT) encompasses a set of mandatory laboratory tests performed before red blood cell transfusion. The antibody screen, one component of PTT, commonly includes a 10-20 min incubation. The primary aim of this study was to determine if this period can be reduced when using current immunohematology methodologies. Antibody screens were performed on reagent samples using Glass or Gel-based column agglutination technologies (CAT) and a solid phase red cell adherence (SPRCA) assay, with incubation periods of 1, 5, 10 and 15 min, and 20 min (SPRCA assay only). For each method, the shortest period producing a minimum of a 1+ reaction with all reagent samples was considered optimal. The sensitivity of each assay using the optimal period was calculated after performing antibody screens on 100 patient samples. It was demonstrated that the incubation period in the SPRCA and Glass CAT systems can be reduced to 5 and 10 min, respectively, while achieving high assay sensitivity (98.9% in both). The incubation period in the Gel CAT system cannot be reduced from 15 min. Significant association between titre and reaction strength was observed for all three screening methods (p < 0.001 for both CAT methods, p = 0.041 for SPRCA). This study demonstrates that the incubation period used in the antibody screen can be reduced when using systems employing the Glass CAT and SPRCA methods, without affecting assay sensitivity. If confirmed, it could result in faster completion of PTT.

Evaluation of Platelet Alloimmunization by Filtration Enzyme-Linked Immunosorbent Assay.

The current methods for detecting antiplatelet antibodies are mostly manual and labor-intensive. A convenient and rapid detection method is required for effectively detecting alloimmunization during platelet transfusion. In our study, to detect antiplatelet antibodies, positive and negative sera of random-donor antiplatelet antibodies were collected after completing a routine solid-phase red cell adherence test (SPRCA). Platelet concentrates from our random volunteer donors were also prepared using the ZZAP method and then used in a faster, significantly less labor-intensive process, a filtration enzyme-linked immunosorbent assay (fELISA), for detecting antibodies against platelet surface antigens. All fELISA chromogen intensities were processed using ImageJ software. By dividing the final chromogen intensity of each test serum with the background chromogen intensity of whole platelets, the reactivity ratios of fELISA can be used to differentiate positive SPRCA sera from negative sera. A sensitivity of 93.9% and a specificity of 93.3% were obtained for 50 μL of sera using fELISA. The area under the ROC curve reached 0.96 when comparing fELISA with the SPRCA test. We have successfully developed a rapid fELISA method for detecting antiplatelet antibodies.

Therapeutic Efficacy of Platelet Transfusion and Analysis of Influencing Factors in 364 patients

Introduction: The aims of this study were to observe the therapeutic effect of platelet (PLT) transfusion and to analyze influencing factors for the sake of providing useful clues for improving the efficacy of PLT transfusion. Methods: Included in this study were patients who received PLT transfusion in the affiliated hospital of Nantong University. Patients’ sex, age, height, weight, PLT transfusion status, and 20–24-h PLT count before and after PLT transfusion were collected to calculate the PLT corrected count increment values before and after PLT transfusion. Solid-phase red cell adherence assay was used to determine PLT antibody. Statistical analysis was performed using SPSS25.0 Software. Results: A total of 364 patients received 1,060 PLT transfusions, including 728 successful transfusions and 332 unsuccessful transfusions. When the patients were grouped according to different etiologies, significant differences in PLT transfusion effectiveness were observed between these groups (χ² = 15.070, p < 0.05). Grouping of the 364 patients according to sex, blood type, and PLT transfusion frequency showed no significant difference in PLT transfusion refractoriness (PTR) between different age-groups and sexes (p > 0.05). With the number of PLT transfusions increasing, PTR increased gradually. PLT antibodies were detected of 364 patients, 67 of them were positive. Among them, 63 cases (94.02%) were positive for HLA class I antibody. Conclusion: To reduce PTR, multiple factors should be considered comprehensively when PLT transfusion therapy is to be implemented in clinical practice. PLT antibody is the main immune factor causing PTR.

Autoantibodies mimicking alloantibodies: A case series unveiling the dilemmas of transfusion.

Autoimmune hemolytic anemia is characterized by increased red cell destruction and/or decreased red cell survival due to autoantibodies directed against self-antigens on red cells. Since autoantibodies react with self and nonself red blood cells (RBCs), they tend to mask the underlying clinically significant alloantibodies and many a times mimic a specific pattern like alloantibodies. We discuss three immune hematological cases of warm autoantibodies. Antibody screening was performed by solid-phase red cell adherence (SPRCA) technique on a fully automated platform NEO Iris (Immucor Inc., USA). In case of a positive antibody screen, antibody identification was performed using SPRCA, NEO Iris (Immucor Inc., USA). Alloadsorption for adsorbing the autoantibodies was done using in-house prepared allogenic packed RBCs - R1R1, R2R2, and rr. All cases had warm autoantibody with a broad specificity against self-Rh antigens. Anti "C" and Anti "e" antibodies were identified in case 1 and autoanti "e" antibody in cases 2 and 3. Case 3 had underlying alloanti "E" along with autoanti "e" which posed a transfusion challenge. Our case series highlights the importance of detecting the nature of the antibody whether it is alloantibody or autoantibody with antigen specificity. This would help in selecting appropriate antigen negative blood units for transfusion purpose.

A rare case of clinically significant naturally occurring anti-Cw reported in a healthy male donor.

Anti-Cw antibody is an immunoglobulin against the red cell antigen Cw which is a low frequency red cell antigen that belongs to the Rh antigen system. It is a clinically significant antibody and may cause haemolysis on exposure to antigen positive red cells. Due to its low frequency, it is not included in routine antibody screening panels. A 32years healthy male donor with no history of transfusion donated whole blood at the department of Transfusion Medicine & Blood Centre of our institute. As a part of routine pre-transfusion testing, the donor's samples were subjected to automated blood grouping and screening for unexpected red cell antibodies using 3 cells panel on solid-phase red-cell adherence (SPRCA) (Galileo Neo, Immucor, Norcross, USA). The antibody screening came out to be positive with a reaction in cell I of the antibody screening panel. Further the antibody was identified as anti Cw in using 16 cells panel, select cells and phenotyping. In the present case, the anti-Cw antibody was found to be reactive at 37°C and AHG phase which could lead to haemolytic transfusion reaction. The fact that the male donor had no history of transfusion or transplant led us to the conclusion that it was a naturally occurring, but a clinically significant antibody. This case highlights the importance of performing an antibody screening for healthy donors as well and urges transfusion services to procure screening cells which incorporate Cw positive cells.

A prospective observational study to compare abo isoagglutinin titer by conventional test tube and column agglutination technique after heat inactivation of plasma in O blood group individuals

Background and Aims: When determining ABO antibody titers, IgG antibodies can be masked by immunoglobulin M (IgM) antibodies. Hence, the measurement of actual concentration of IgG requires methods such as heat inactivation (HI) of plasma. This study was aimed at determining the effects of HI on IgM and IgG titers performed by conventional test tube (CTT) technique and column agglutination technique (CAT). Materials and Methods: This was a prospective, observational study conducted from October 2018 to March 2020. All consecutive O group donors who gave consent for participation were included in the study. All samples were consecutively tested by CTT and CAT, before and after HI (pCTT and pCAT). Results: A total of 2005 donors were included. IgG titers were found to be more than IgM titers. pCTT IgG results are similar or lower when compared to results obtained by solid-phase red cell adherence (SPRCA), whereas pCAT IgG titers were higher with pCAT when compared to hemagglutination (HA)/SPRCA. The results of titers obtained by pCTT were lower as compared to pCAT and HA/SPRCA, with majority giving results <64. The median IgG and IgM titers for both anti-A and anti-B were highest in pCAT, whereas the median IgG and IgM anti-A and anti-B titers were similar to HA and pCTT. Conclusion: Results obtained by HA/SPRCA were closer to results obtained by pCTT, with the advantage of less time consumption, automation requiring less expertise, and no interobserver variation. Titers obtained by pCAT were higher in comparison to HA/SPRCA and pCTT results, due to high sensitivity.

Investigation of Apparent Solid Phase-Only Red Cell Serologic Reactivity: Clinical and Laboratory Correlations.

Solid phase red cell adherence is a sensitive platform for blood group antibody detection, but non-specific reactions may occur. The aim of this study was to define the clinical characteristics of patients with these reactions and their associated laboratory findings. An 8-month retrospective review of a regional blood bank database was performed. One hundred and seventy-three patients were identified with apparent nonspecific solid phase (NSP) reactivity. Serologic findings were recorded, and each patient's electronic health record was interrogated. NSP reactivity was the most common positive finding in the laboratory. Of 173 patients with NSP, 167 had documented concurrent tube testing. Of these, 165 were negative, one demonstrated nonspecific reactivity, and one anti-Lea was identified. Most positive solid phase antibody screens had negative panel testing, with fewer cases of pan-reactivity or sporadic reactivity. Follow-up testing was either negative (85.5%) or demonstrated NSP reactivity (14.5%). No new blood group antibodies were identified. Most patients were female (72.8%), with pregnancy being the leading diagnosis (35.8%); however, this mirrored the distribution of cases received in the laboratory. When pregnant patients were excluded, female and male patients had the same average age, and the gender distribution and the primary diagnoses of NSP patients mirrored those of all patients evaluated. Solid phase antibody detection is known to be sensitive, but nonspecific reactions are relatively common. Compared to other studies, the evolution of NSP to clinically significant antibodies was not seen, female patients did not have a predilection for NSP reactivity, and NSP was not associated with certain diagnoses.

Evaluation of the effectiveness of platelet crossmatching by the solid-phase red cell adherence assayin adult patients of a tertiary care hospital in Thailand: A retrospective study.

Background and AimsPlatelet transfusion refractoriness is well aware to be associated with poor clinical outcomes. Patients with the alloantibody causing refractoriness required cross‐matched compatible products to improve the platelet number. This study aims to evaluate the effectiveness and availability of platelet crossmatching provided by the solid‐phase red cell adherence (SPRCA) technique in the context of a tertiary university hospital.MethodsA retrospective chart review was performed of the records of 214 patients with platelet refractoriness in Siriraj Hospital, a tertiary university hospital in Thailand, between January 1, 2017, and December 31, 2020.ResultsThe SPRCA technique successfully provided cross‐matched compatible platelets to 114 patients (69.7%). Platelet crossmatching significantly improved the platelet counts, as shown by the increased 1‐ and 24‐h corrected‐count increments (p< 0.0001). No acute transfusion reactions were observed in these patients. Of the 114 patients who received cross‐matched platelets, 82 patients (71.9%) survived at 30‐day posttransfusion; whereas, 16 patients (14.0%) died within 7‐day posttransfusion.ConclusionThe SPRCA method can provide a high availability rate of cross‐matched platelets, which is effective at stopping and preventing clinical bleeding conditions. This method is appropriate to apply for platelet crossmatching in the context of a hospital blood bank.

Evaluation of Human Leucocyte Antigen Mediated Platelet Transfusion Refractoriness and Platelet Crossmatching Assay in Patients with Hematologic Disorders.

ObjectivesPlatelet refractoriness complicates the platelet transfusion, which is essential for managing thrombocytopenia in patients with hematological disorders. It is associated with adverse clinical outcomes and increased health care costs. We conducted a prospective study to determine the effectiveness of cross-matched compatible platelets in a group of patients refractory to platelets from random donors and to evaluate human leukocyte antigen (HLA)-mediated refractoriness.MethodsThis prospective study was conducted on 40 patients with different hematological disorders requiring platelet transfusions who were refractory to random platelets and presented to the hematology unit of Alexandria's main university hospitals between May 2020 and March 2021. They received 60 ABO-compatible platelet transfusions, either leuco-reduced or random donor platelets, stored for no more than 72 hours. A solid-phase red cell adherence technique (SPRCA) was used for platelet crossmatching. The corrected count increment (CCI) was used to monitor the effectiveness of each platelet transfusion with a cut-off value of 5 × 103/µL at 1 hour and 2.5 × 103/µL at 24 hours. Anti-HLA antibodies were assessed using the enzyme-linked immunosorbent assay technique.ResultsOut of 60 cross matches, 47 (78.3%) were compatible, and 13 (21.7%) were incompatible. Among 47 compatible results, 30 (63.8%) showed adequate CCI and 17 (36.2%) showed inadequate CCI at 1-hour post-transfusion. Among the incompatible results, 3 (23.1%) had adequate CCI and 10 (76.9%) had inadequate CCI. Significant improvements were found in the mean CCI when comparing cross-matched compatible platelets and incompatible platelets at 1hour or 24 hours (p=0.009 and p < 0.001, respectively). From the 40 studied patients, HLA alloimmunization was present in 18 patients (45.0%) and absent in the remaining 22 patients (55.0%). In the absence of HLA alloimmunization, patients showed significantly better responses at 1 hour and 24 hours (p =0.001 and p =0.015, respectively). There was better sensitivity of platelet crossmatching with random donor platelet concentrates than single donor platelet concentrates.ConclusionsPlatelet crossmatching using SPRCA and HLA screening are effective and rapid tools for better management of patients' refractory to platelet transfusions.

Comparative Evaluation of heat-inactivated ABO isoagglutinin titers with solid-phase red cell adhesion titers

Background and Objectives: Measurement of actual concentration of IgG requires methods like heat inactivation (HI) of serum. This study was aimed at comparing of heat-treated ABO titers performed by conventional test tube technique (CTT) and column agglutination technique (CAT) with hemagglutination (HA)/solid-phase red cell adherence (SPRCA). Methods: This was a prospective, observational study conducted between October 2019 and March 2020. All consecutive A, B, and O group donors who gave consent for participation in the project were included. All samples were tested by CTT and CAT before and after HI (post DTT treatment performed by CTT [pCTT] and post DTT treatment performed by CAT [pCAT]) and with HA/SPRCA. Results: A total of 300 donors were included: 100 each with A, B, and O blood groups. For group O, IgG titers were higher than IgM titers among group O, whereas that among group A and group B, IgM titer was found to be higher than IgG titer by both HA/SPRCA and pCTT. The titer values among all the three blood groups were highest when the test was performed by pCAT, though the results of pCTT and SPRCA, pCTT, and pCAT were significant in group O donors. For groups A and B, results of pCTT and pCAT, pCAT, and HA/SPRCA were found to be statistically significant. Conclusion: Results obtained by HA/SPRCA were closer to pCTT, with respect to time consumption, expertise required, and lack of inter-observer variation. Titers obtained by pCAT were higher in comparison to pCTT obviously due to its relative higher sensitivity. Hence, these methods cannot be used interchangeably.

Evaluation of influence of IL-6 C-572G gene polymorphism and clinical factors on positive platelet antibody test

Background: Interleukin-6 (IL-6) promotes antibody production. The objective of this study was to investigate whether IL-6 C-572G single nucleotide polymorphisms (SNP) and clinical factors are associated with positive platelet antibody test. Materials and methods: Thirty platelet recipients with platelet antibodies (responders) and 20 platelet recipients without platelet antibodies (non-responders) were randomly selected. The -572 C&gt;G (rs 1800796) SNPs in the promoter region of IL-6 gene were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Solid phase red cell adherence assay (SPRCA) was used for platelet antibody detection. Results: Age, sex, percentage patients with benign diseases, and percentage of patients with homozygotes for the C allele at position -572 of the IL-6 gene were similar between responders and non-responders. Although the amounts of platelets pheresis transfused to patients with hematologic diseases were higher than those of non-hematologic diseases (47.2 ± 54.2 vs. 17.4 ± 13.8 units, p = 0.019), detection rate of platelet antibodies was lower in patients with hematologic diseases than that in patients with non-hematologic diseases (42.3% vs. 79.2%, p = 0.01). Conclusion: There was no association between IL-6 C-572G gene polymorphism and positive reactivity in solid phase platelet antibody detection method in platelet recipients.

Application of lyophilised human platelets for antibody detection in solid phase red cell adherence assay

Antibodies against human platelets cause a variety of thrombocytopenic disorders, which lead to potentially fatal haemorrhage. Therefore, their prompt detection is mandatory for successful patient treatment. Solid phase red cell adherence (SPRCA) assay allows for platelet antibody detection widely. However, preparation of fresh platelets with HLA-I and human platelet antigens (HPA)1–5,15 genotyped as target cells is inconvenient and fresh platelets have a short shelf life. In this study, the lyophilised human platelets for antibody detection in SPRCA were prepared. Firstly, platelets were resuspended in lyophilisation buffer and freeze-dried. Then the characteristics of lyophilised platelet were analysed. Rehydrated platelets were recovered with a mean rate of 80.91% ± 2.87%, and still retained spherical morphology. Indirect flow cytometry showed that glycoproteins IIb/IIIa, Ia/IIa, Ib/IX, IV, CD109, and HLA class I were present on the surface of the lyophilised platelets at a comparable level to that of fresh platelets. The consistent results obtained with WHO reference reagents containing anti-HPA-1a, anti-HPA-3a, and anti-HPA-5b, as well as clinical samples from the same donors containing anti-HLA antibodies when reacting with lyophilised versus fresh platelets confirmed good antigenicity preservation of platelets after freeze-drying. Further investigation showed that the lyophilised platelets could be stored at 2–8 °C for up to 14 months and the reconstituted suspension was stable for 48 h. Therefore, lyophilised platelets can be a convenient alternative to fresh platelets to use for anti-platelet antibody detection in SPRCA tests.

Correlation of platelet crossmatch results by Solid Phase Red Cell Adherence Assay (SPRCA) with post-transfusion platelet count increment in adult hemato-oncology patients of a tertiary care oncology centre in India

AimsTo assess platelet crossmatch result by SPRCA and find its correlation with post-transfusion platelet count increment among adult hemato-oncology patients. MethodsA prospective observational pilot study of 50 adult hematologic malignancy patients previously transfused, but not already known to be transfusion-refractory and without any nonimmune causes for inadequate response to platelet transfusion were included after obtaining informed consent. They were transfused one unit of ABO identical single donor platelet. Ten minutes to 1 -h post-transfusion CCI was calculated.CCI ≥ 7500 was considered as adequate response. Post-transfusion crossmatching by SPRCA was performed by using preserved platelet samples from donor units with the serum of the respective patient. Statistical analysis of the correlation between platelet crossmatch results and CCI was done. ResultsOut of 50 crossmatches, 78% (39/50) showed compatible and 22% (11/50) showed incompatible results. Among 39 compatible results, 87.2% (34/39) showed adequate CCI and 12.8% (5/39) showed inadequate CCI. Among 11 incompatible results, 18.2% had adequate CCI and 81.8% had inadequate CCI. The difference between the response in terms of CCI to compatible and incompatible crossmatches was found to be statistically significant (p < 0.05). Other variables like age, sex, number of previous transfusions and underlying clinical condition of the patient were not found to have any effect on the compatibility of crossmatch. ConclusionsTransfusion of crossmatched platelets to non-refractory, multiply transfused hematological malignancy patients without serious illness might provide a small benefit over transfusing randomly selected platelets, though these data must be confirmed with a larger sample size.

Efficacy of cross-match compatible platelets in multi transfused haemato-oncology patients refractory to platelet transfusion

BackgroundPlatelet refractoriness, which leads to platelet transfusion failure resulting in significant morbidity and long hospital stay, is routinely not investigated. AimsTo determine the efficacy of cross-match compatible platelets in multi-transfused alloimmunized hemato-oncological patients refractory to platelet transfusion. Materials and method149 ABO compatible single donor apheresis platelet transfusions given to 38 alloimmunized refractory patients. Corrected Count Increment (CCI) <5000 (1 h) was taken to define refractoriness. Solid-phase red cell adherence assay was used to determine the alloimmunization status and platelet cross-matching. Post Transfusion Platelet Increment, CCI and the Percentage Platelet Recovery were used to monitor the effectiveness of platelet transfusion. ANOVA test followed by Post hoc test Tukey HSD used to compare group means and classified into three groups depending upon the cross-matching and compatibility status. Categorical data was analysed for various outcomes using Pearson's chi square test or Fischer exact test. ResultPatients showed statistically significant recovery in terms of PPI, CCI and PPR at 1 h post SDAP transfusions when they received cross-matched compatible platelets. The one-hour CCI was significantly higher for cross-match-compatible platelets (19173 ± 2692) than for incompatible (5888 ± 1526) and for uncross-matched (8140 ± 1480). Forty four (97.8%) of 45 cross-matched compatible platelet transfusion episodes showed a satisfactory response in terms of PPI and CCI values as compared to 50 % and 53.9% in uncross-matched group respectively (p < 0.0001). ConclusionPlatelet cross-matching is an effective intervention in the management of multi-transfused alloimmunized Haemato-oncological patients, refractory to platelet transfusion.